Scaffolding proteins altered in the ability to perform a conformational switch confer dominant lethal assembly defects

James E. Cherwa, Asako Uchiyama, Bentley A. Fane

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

In the φX174 procapsid crystal structure, 240 external scaffolding protein D subunits form 60 pairs of asymmetric dimers, D1D 2 and D3D4, in a non-quasi-equivalent structure. To achieve this arrangement, α-helix 3 assumes two different conformations: (i) kinked 30° at glycine residue 61 in subunits D 1 and D3 and (ii) straight in subunits D2 and D4. Substitutions for G61 may inhibit viral assembly by preventing the protein from achieving its fully kinked conformation while still allowing it to interact with other scaffolding and structural proteins. Mutations designed to inhibit conformational switching in α-helix 3 were introduced into a cloned gene, and expression was demonstrated to inhibit wild-type morphogenesis. The severity of inhibition appears to be related to the size of the substituted amino acid. For infections in which only the mutant protein is present, morphogenesis does not proceed past the first step that requires the wild-type external scaffolding protein. Thus, mutant subunits alone appear to have little or no morphogenetic function. In contrast, assembly in the presence of wild-type and mutant subunits is blocked prematurely, before D protein is required in a wild-type infection, or channeled into an off-pathway reaction. These data suggest that the wild-type protein transports the inhibitory protein to the pathway. Viruses resistant to the lethal dominant proteins were isolated, and mutations were mapped to the coat and internal scaffolding proteins. The affected amino acids cluster in the atomic structure and may act to exclude mutant subunits from occupying particular positions atop pentamers of the viral coat protein.

Original languageEnglish (US)
Pages (from-to)5774-5780
Number of pages7
JournalJournal of virology
Volume82
Issue number12
DOIs
StatePublished - Jun 1 2008

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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